AAV9载体在小鼠体内分布的长期研究

Long-term Study on the Distribution of AAV9 Vector in Mice

目的研究AAV9载体在小鼠体内的长期分布情况,从而为CRISPR/Cas9技术用于治疗DMD患者的安全有效性提供研究基础。方法将实验小鼠分为A、B两组,A为实验组、B为对照组,在处理后的第1、3、7、14、30天观察小鼠体内的荧光表达情况并对其主要肌肉组织进行荧光定量分析。结果经肌肉向小鼠体内注射AAV9-Luc后,在30天内,随着时间的延长,荧光表达量逐渐增加,表达先局限于注射处,后渐渐扩大,且未见除肌肉外其他器官有明显荧光表达。结论AAV9载体在小鼠体内局限分布于肌肉组织,而在非肌肉器官中几乎没有分布,具有很高的肌肉组织特异性。初步证明在AAV9载体介导CRISPR/Cas9治疗DMD技术中,经肌内注射的安全性较高。

Objective To investigate the long-term distribution of AAV9 vectors in mice to provide a basis for research on the safety and efficacy of CRISPR/Cas9 technology for treating patients with DMD.Methods The experimental mice were divided into two groups,A and B,with A being the experimental group and B being the control group.The expression of fluorescence in the muscles of the mice was observed on days 1,3,7,14 and 30 after treatment and the virus in them was quantified.Results After the intramuscular injection of AAV9-Luc into mice via muscle,the fluorescence expression gradually increased with time over 30days,and the expression was first confined to the injection site and then gradually expanded,and no significant fluorescence expression was seen in organs other than muscle.Conclusion AAV9 vectors are restricted to muscle tissues in mice and have little distribution in non-muscle organs,with high muscle tissue specificity.It was preliminarily proved that the intramuscular injection of AAV9 vector-mediated CRISPR/Cas9 in DMD was safe.